Apparatus for non-invasive forced oxygen enrichment of avian eggs

ABSTRACT

Apparatus, methods and products in which an egg is non-invasively infused with oxygen such that the structural integrity of the shell of the egg is not compromised. The method comprises the step of subjecting the external surface of the egg shell to the oxygen under a condition of at least one of vacuum and positive pressure. The oxygen is from a substance selected from the group consisting of oxygen, nascent oxygen, liquid oxygen, oxone, ozone, hydrogen peroxide, and potassium permanganate.

BACKGROUND OF THE INVENTION

In non-extinct species, the avian egg is known to range in mass from0.25 to 1,500 grams. The common poultry egg has an average weight ofabout 60 grams. The common fertilized poultry egg is both self-containedlife support system and cradle. However, the life support system isflawed. It lacks oxygen. All other elements are present, including thefuel with which the fire of life will be sustained through thetwenty-one day journey.

Oxygen is the driving force which feeds the biological matrix from whichthe new life is fashioned. The cycle of synthesis begins at the moment"set" occurs, i.e. the moment that the process of viability is initiatedwhen the temperature of the fertilized egg exceeds about 68° F. to about70° F. for any substantial period of time.

From the first moment the process of viability begins, the margins ofsuccess are tenuous. A race between catabolic and anabolic processes isinitiated as life force courses forward. Oxygen must be obtained and theapparatus for gathering it must be fashioned within the margins of thesmall reserves available. Never again will the challenge be quite sogreat or the margins so small. The slightest unfavorable environmentalconditions, and the battle is lost.

Unfortunately, the optimal conditions for incubation also favor internalcompetitive breakdown forces and external invasive ones. Idealconditions in the incubator, darkness, moderately high humidity andtemperatures of 99.5° F.±0.5° F. are nearly optimal for microbial lifewhich will proliferate around and inside the egg shell. Thus, amultitude of foreign life forms are present. Some may be innocuous,others competitors or antagonists, and it is even possible that somefamiliar types such as Salmonella are symbiotic to the avian neonate.

The viability processes begin at the center but are subsequently focusedoutward to the extremities of the egg. Surrounding the embryo is alaminar set of membranes. The outermost layer (actually, a bi-layer)lies against the shell. The innermost layer is referred to as thechorioallantoic membrane and envelopes the albumen and yolk.

The outer shell is made up of crystalline calcium and is many timesthicker than the inner shell and other adjacent membranes combined. Theexternal surface has trumpet-shaped pores which run through the shell tothe surface to the membrane. Typically, there are about 7,000 to 17,000pores distributed over the shell surface, the highest density being atthe rounded end (top) of the egg proximal to the air sac locatedadjacent to the interior of the shell. The air sac is a space created bya parting between the outer and inner shell membranes wherein the outermembrane remains attached and the inner (chorioallantoic) membrane isdetached from the outer shell membrane. The separation between the twomembranes creates the air sac which serves as a gas exchange depot,sometimes referred to as the "dumb" lung. The dumb lung respires inresponse to changes in temperature and atmospheric pressures. The dumblung also grows larger and larger as the ovum takes up less and lessspace in using up its reserves, the byproducts of which pass through thelung to the outside. Once incubation has started, the embryo provides anetwork of arteries and veins to the working side of the lung, whichthereafter allows for efficient gas exchange through the lung.

Prior art teaches that as pressure is applied and then increased tofertilized eggs, fatalities from the onset of pressure very quicklyincrease to 100%. Prior art also teaches that as vacuum is applied andthen increased to fertilized eggs, fatalities from the onset of pressurevery quickly increase to 100%. Furthermore, prior art shows that whenambient oxygen is increased or decreased above normal concentrations(21%) during incubation, the hatch rate of fertilized eggs decreasesmarkedly. In sum, prior art shows that there is no known advantage toany process applied to fertilized eggs prior to incubation, and thatfertilized eggs are adversely affected by such treatments.

Handling and artificial incubation methods have developed over the lastcentury to create the best known conditions of humidity, aircirculation, stable storage and incubation temperatures, and motion toprovide maximum hatch of fertilized egg stock. For storage of fertilizedegg stock, air sac (top) up for no longer than twenty days at atemperature of about 55° F. is preferred. For incubation, air sac (top)up, humidity of about 60% during the first 18 days and about of 70% thelast three days, a temperature of 99.5° F. ±0.5° F. with a gentle,cradle-like rocking motion, timed about once every ninety minutes, andfiltered, recirculated ambient air (21% O₂) is preferred. Theseconditions result in more or less maximum average hatches. Percentagesof average hatches vary considerably depending upon the storage age offertilized egg stock and the age, type and conditions of laying flocksfrom which they are derived. In general, the range is from about 80% tounder 86% hatch, or a 14% to 20% mortality less the percentage ofunfertilized eggs.

Shortly after hatching, chicks are injected with vaccines (such asMarek's) to prevent flock diseases. Significant expense and mortalityare incurred due to the invasive nature of the injection.

Improvements in the overall healthiness of the hatched chicks stronglyinfluences efficiencies with respect to grow-out factors of the flock,i.e., percentages of grow-out mortality, size, strength, fertility, feedconversion, susceptibility to disease and, ultimately, overallcommercial success of the flock as layers, breeders or broilers andfryers for meat. Improvements related to better disease resistance, feedconversion, strength and increased commercial success from grow-out arealso desirable. Finally, the ability to provide early nutritionaladvantages, to provide medications related to improved mortality orgrowth, to alter or manipulate sex characteristics, to provide microbialsynergists (probiotics), to control disease agents, including those ofpoultry and those pathogenic to humans, is also desirable.

SUMMARY OF THE INVENTION

Oxygen as defined herein includes O (oxone), O₂, O₃ (ozone) or anyconvenient source thereof including liquid oxygen, hydrogen peroxide andpotassium permanganate, all of said chemicals above defined asoxygen-bearing chemicals.

Fertilized egg as defined herein includes both fertilized andpre-fertilized eggs (ovum)--also referred to as "vital eggs"--of anybird egg including chicken, turkey, goose, duck, raptor and parrot.

The present invention encompasses treating fertilized eggs prior to andin prenatal incubation phases by methods employing one or more ofcombinations of pressures, vacuums and above ambient levels of oxygen inthe form of O₂ and/or O₃, or sources thereof, to non-invasively transferthe O₂ and/or O₃ through the egg shell without compromising thestructural integrity of the shell.

When used according to the present invention to achieve the purposes asoutlined, ozone exhibits vitalizing, as opposed to expected lethalproperties, endowing the ovum with remarkably improved strength. Thisnot only results in significantly higher survivability throughincubation, but also greater strength through adulthood.

Contrary to the prior art, it is in the period of incubation and beforethe egg has developed oxygen-collecting organs or respiration thatforced oxygen/ozone treatment demonstrates the most profound overallresults. Increased hatch of fertilized eggs may be obtained by oxygenenrichment, preferably forced, and most preferably when the treatment isapplied prior to or soon after incubation starts, with the oxygen beingsupplied into the egg spaces and interstices--even in high concentrationlevels which would normally be considered lethal concentrations.Non-invasive oxygen augmentation may be accomplished in a variety ofways, including subjecting fertilized eggs to brief periods of vacuumsand/or pressures in the presence of oxygen or oxygen-containingsubstances or carriers.

Non-invasive but forced perfusion of disease control vaccines offertilized eggs, with or without simultaneous forced enrichment ofoxygen, can also provide profound benefits in terms of reduced mortalityand cost savings of inoculation. More certain inoculation andsubstantial reductions in hatchling injection mortality occur.

Additionally, non-invasive forced perfusion of growth, health and gendercharacteristic additives, such as aromatase, may be added with orwithout enriched oxygen, affording valuable benefits to treated hatchedflocks including important "out of the shell" disease and infectioncontrol, improved growth, feed conversions and added overall robustnessand vitality.

Moreover, the levels of enrichment required to improve survivability offertilized eggs also endow the chicks with greater robustness, as shownby better feed conversions over the grow-out period. The result is muchhigher efficiencies of production. Feed conversion is a composite resultwhich may be comprised of one or more other operative factors such ashealthier, stronger, less diseased or more disease resistant chicks.

Also, the microbicidal effects of oxygen against poultry endemicbacteria, such as Salmonella, are realized; particularly if more activeforms such as nascent and ozone oxygen are increased in the oxygenenrichment treatment. Finally, the present invention provides improvedfood safety, for it is axiomatic that healthier animals providehealthier, safer food products.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing objects and attendant advantages of this invention willbecome more readily appreciated as the same becomes better understood byreference to the following detailed description when taken inconjunction with the accompanying drawings:

FIG. 1 is a block diagram of a first exemplary apparatus of the presentinvention;

FIG. 2 is a block diagram of a second exemplary apparatus of the presentinvention;

FIG. 3 is a block diagram of a third exemplary apparatus of the presentinvention;

FIG. 4 is a block diagram of a fourth exemplary apparatus of the presentinvention;

FIGS. 5A-5C are schematic representations of a non-invasive localizedoxygen/ozone augmenting apparatus of the present invention;

FIG. 6 is a schematic representation of a non-invasive systemicoxygen/ozone augmenting apparatus of the present invention;

FIGS. 7A-7H are schematic representations of a second non-invasivelocalized oxygen/ozone augmenting process and apparatus of the presentinvention;

FIG. 8 is a graphical representation of the vacuum and pressureprocesses of the present invention;

FIG. 9 is a graphical representation of the vacuum process of thepresent invention;

FIG. 10 is a graphical representation of the pressure process of thepresent invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS I. OVERVIEW

A. Oxygen.

Application of oxygen may be calculated as any equivalent mole weightabove atmospheric concentrations (20.946%) supplied to the fertilizedegg. Oxygen includes allotropic forms, nascent oxygen, liquid oxygen andoxygen-containing compositions such as hydrogen peroxide (H₂ O₂),potassium permanganate (KMnO₄), and the like. Mole weights, calculatedas percentages of the oxygen in enrichment fluid supplied to thefertilized egg, may be calculated as O₁ (oxone) at mole weight 7.9997,O₂ (oxygen) at mole weight 15.9994, and O₃ (ozone) at mole weight47.9982.

For purposes of increased hatchability of fertilized eggs, the workingrange of enrichment oxygen calculated as O₂, O₃ or a combination thereofwhich may be employed is from greater than ambient, and is from about21% to about 99.9998%. The preferred range of oxygen is from about 25%to about 99.99%. The most preferred percentage of oxygen is 45%±4%. Whenreduction of indigenous microbes is the object, or an added object, thepreferred range is from about 50% to about 95%, and the most preferredoxygen concentration is about 70% ±5%.

For hatchability, the oxygen may be high purity oxygen or a mixture ofhigh purity oxygen (O₂) and ozone (O₃) as liquids or gases. Oxygensources may also be selected as hydrogen peroxide, potassiumpermanganate solution and other fluidized oxygen carriers. A mixture ofO₂ and O₃ are preferred, from about 1 to about 250 parts per million ofO₃ to O₂.

For hatchability and microbial control, a mixture of O₂ and O₃ may alsobe used, with O₃ in the range of about 10 to about 5,000 parts permillion of O₂. A ratio of about 100 parts per million O₃ to O₂ ispreferred. Though not preferred, H₂ O₂, O₂ and O₃ may also be used incombination, or H₂ O₂ may be used as an adjunct or treating agent inranges of from about 0.001% to about 30% in aqueous solution. About7%±about 4% is preferred. Aqueous solutions of potassium permanganatemay range from about 0.001% to about 20% in aqueous solution, but arepreferred in lower concentrations, i.e., about 3%±about 2%.

Oxygen working range by weight compared to weight of egg is about 0.0001grams to about 20 grams per 1,000 grams of egg mass. As stated, theoxygen may be from any source, such as liquid oxygen, purified oxygen,oxygen or ozone dissolved or associated with water or other carrier(s)or solute(s), including but not limited to ambient air as an ozonecarrier. The preferred range is from about 0.001 to about 9 grams per1,000 grams of egg mass. The preferred amount is about 3.0 grams per1,000 grams of egg mass. Where oxygen or an oxygen source alone is usedas the treating agent or in a carrier (usually but not always one orboth of air or water), the concentrations may range from more thanambient (21%) to about 99.9998%. The preferred range is from about 60%to about 99%. Some other preferred levels of oxygen as treating agent oringredient thereof are: when applied as a gas including available oxygenin carrier, about 95%; when applied as a liquid with carrier(s), about50%; when applied as a super-cooled or liquid gas, about 99.8%; whenapplied as a super-cooled liquid or gas containing at least 1 part permillion ozone, about 99%; when applied as hydrogen peroxide solution,about 60%; and as hydrogen peroxide and/or permanganate solutioncontaining at least 1 part per million ozone, about 70%. Ozonatedmixtures of oxygen are the most preferred enrichment media.

B. Negative Pressure:

Fertilized eggs may be subjected to negative pressure at the externalsurface of the egg. One method is to place the egg in an enclosed plenumupon which is exerted negative pressure to create a vacuum. Vacuums areapplied externally to the egg in order to create room within the egg forthe oxygen, and to force enriched forms of oxygen(s) and/oroxygen-carrying substance(s) through the shell upon relief of negativepressure. Enrichment capacity of the egg is enhanced by disinfusing thevacuum of ambient gases previously present in internal spaces and asdissolved gases in the egg matter. This step is followed by judiciousintroduction of the selected oxygen enrichment mixtures into thevacuumized plenum and consequently into the egg, which is also in astate of negative pressure. Alternative methods of treatment may also beused, though the underlying principles remain operative.

The above method of treatment permits optimization of the maximumconcentration of selected enrichment gases into the egg. Such means canbe accomplished by pathways other than using a chamber or vessel. Forexample, the egg may be warmed to cause internal expansion, resulting inexpelling of ambient internal gases, and then cooled in the presence ofselected enrichment gases, whereupon the egg will take in enrichmentgases and replace at least some of the original ambient gases present.Also, liquid oxygen, preferably containing ozone, may be applied by finespray to the pores of the egg surface to infuse oxygen. Another exampleis to employ a pressure cup, such as a suction cup, which can be affixedby contact to the external shell surface, preferably immediately overthe air sac where poration is at maximum density. By applying negativepressure vacuum through an aperture in the suction cup, the air sac andother portions of the egg are emptied of indigenous gases. Oxygen andother treating agents may then be infused through the shell pores bynegative pressure and/or in a positive pressure stream. While notessential to minimal oxygen enrichment, treatment vacuums are preferredwhen gasses alone are employed. An exception to the use of vacuum iswhere the available oxygen or other treating agent is in a liquid orassociated with a carrier, such as liquid oxygen, hydrogen peroxidesolutions or potassium permanganate in aqueous solution, as examples. Inthese instances, a positively pressurized fluid stream containingperfusate alone may be directed across and forced through the poresadequately. However, as a rule both negative and then positive pressureare preferred steps.

The working ranges of vacuums which may be employed are from greaterthan ambient to about 29.99 inches of mercury for a short time period.The time range for application of preferred vacuums from ambient toabout 29.50 inches of mercury is from about one second to twentyminutes. The preferred vacuum range is from about 2.00 inches of mercuryto about 29.99 inches of mercury. The preferred time range forapplication of the above preferred range of vacuum is from about one toabout ten minutes. The preferred vacuum, if no added pressure cycle isemployed, is about 28.00 inches of mercury ±1.5 inches of mercury. Thepreferred time at preferred vacuum, if no added pressure cycle treatmentis employed, is about one and one half minutes ±one minute. If apositive pressure cycle also is to be applied to the egg, then thepreferred vacuum is still about 28.00 inches of mercury ±1.75 inches ofmercury for about one and one half minutes ±one minute.

Generally, the time to achieve the elected vacuum may range from aboutone second to twenty minutes. Preferred times to achieve elected vacuumsfor all preferred ranges is from about thirty seconds to about tenminutes. Preferred time to achieve vacuum is about three minutes ±1.5minutes. If a given vacuum is achieved too quickly, such as 29.90 inchesof mercury in one second, physical damage can occur to some eggs. Theopposite is true for the time over which the vacuum is held. If held toolong at higher vacuum ranges, e.g. above about 27 inches of mercury fortwelve to thirty minutes, the unabated vacuum may cause damage to someeggs.

Negative pressure may not be required for lower efficiency treatment;positive pressure alone may be adequate. Of course, the reverse is alsotrue. Where lower efficiency treatment is acceptable, however, positivepressure is usually preferred since it alone fills the air sac. All butthe most modest (approximately three inches of mercury to sixteen inchesof mercury) negative pressure treatment will collapse the air sac andthe air sac can only be refilled by at least a small (approximately fiveto ten pounds per square inch in a plenum) positive pressure step.

C. Positive Pressure.

Positive as well as negative pressure without oxygen kills fertilizedeggs. If positive and/or negative pressure is applied for a sufficienttime period or at high or low enough pressure, even added oxygens willnot prevent egg fatalities. The working range of positive pressureswhich may be employed are from slightly greater than ambient (about 3pounds per square inch) to about sixty pounds per square inch for fromabout one second to about thirty minutes. The preferred range ofpressure is from about two pounds per square inch to about thirty-fivepounds per square inch for from about one second to about ten minutes.The preferred pressure and time is about fourteen pounds per square inch±three pounds per square inch for about one and one half minutes ±oneminute. The preferred pressure ranges when a vacuum is also employed arefrom about three pounds per square inch to about twenty pounds persquare inch for from about four minutes to about fifteen minutes, butranges of from about two pounds per square inch to about thirty-fivepounds per square inch from about one second to about ten minutes areacceptable. Preferred pressure with vacuum is about fourteen pounds persquare inch for about two minutes. Working ranges for pressure whenapplied to an oxygen-bearing fluid stream are from about twenty fivepounds per square inch to about 150 pounds per square inch, and whenapplied to fluid stream bursts from about 100 pounds per square inch toabout 1,500 pound per square inch.

II. SPECIFIC EXAMPLES EXAMPLE 1

Referring to FIGS. 1 and 8, fertilized eggs were force-enriched withoxygen as follows. Five hundred twenty eggs were treated and fivehundred twenty were kept as controls. The average weight of the eggs was60 grams±5 grams. Sixty-five eggs were in each treatment, and the totalnumber of treatments was eight. Bottled oxygen (99.8%) waspressure-regulated for delivery to the treatment vessel at 32 pounds persquare inch through a U.V. ozone generator (7.5 watt, 2,537 Å bulb) andU.V. chamber (7-inch diameter×14-inch length). The ozone generated fromoxygen was 0.5 grams/cubic meter of oxygen at 32 pounds per square inch.Sixty-five eggs were placed in the pressure vessel and sealed. Thevacuum level was selected to be 28.5 inches of mercury with elapsed timeto full vacuum being 2.5 minutes. At 28.5 inches of mercury, evacuationof the chamber was stopped and the eggs were held for five minutes. Theequilibrated oxygen in the U.V. chamber at thirty-two pounds per squareinch was slowly fed into the egg vessel to restore ambient pressure. TheO₂ /O₃ continued to be fed into the chamber for about ten minutes untilthirty-two pounds per square inch was achieved. The thirty-two poundsper square inch was held for five minutes. Immediately thereafter, theexhaust valve of the pressure vessel containing the eggs was opened andthe oxygen and ozone were exhausted until the chamber achieved ambientpressure. The time from the exhaust of pressured oxygen/ozone to ambientpressure was approximately one minute. The eggs were removed from thechamber and the process repeated seven times until five hundred twentyeggs had been treated per test. After removal from the chamber, the timefrom the process until the eggs "set" in the incubator ranged from fiveto one hundred twenty minutes. Temperature throughout the process andpreincubation hold time was 60° F.±5° F.

The results were as follows:

    ______________________________________                                        Increase     Hatched  Unhatched   %    %                                      ______________________________________                                        Control Group                                                                              435      85          83.7                                        Group #1     458      62          88.1 5.3                                    Control Group                                                                              417      103         80.2                                        Group #2     437      83          84.0 4.7                                    Control Group                                                                              444      76          85.4                                        Group #3     463      57          89.0 4.2                                    Control Group                                                                              437      83          84.0                                        Group #4     467      53          89.8 6.9                                    Control Group                                                                              433      87          83.3                                        Group #5     456      64          87.7 5.3                                    Control Group                                                                              470      50          90.4                                        Group #6     489      31          94.0 4.0                                    Control Group                                                                              422      98          81.2                                        Group #7     462      58          88.8 9.4                                    Control Group                                                                              450      70          86.5                                        Group #8     471      49          90.6 4.7                                    Control Group                                                                              433      87          83.3                                        Group #9     457      63          87.9 5.5                                    Control Group                                                                              434      86          83.5                                        Group #10    456      62          87.7 5.0                                    ______________________________________                                    

The average increase in hatch for all ten groups was 5.5%.

    ______________________________________                                        Grow-Out Test                                                                            Hatched  Unhatched  %    % Increase                                ______________________________________                                        Control Group                                                                            435      85         83.7                                           Group #11  458      62         88.1 5.3                                       ______________________________________                                    

After hatch, both the treated and control chicks from Group No. 11 werethen transported to a grow-out facility where feed consumption and bodyweight were monitored weekly.

Feed consumption is typically calculated based on the amount of feednecessary to increase the bird weight by one pound. This is referred toin the industry as the feed conversion ratio. Efficiencies in feedinghave increased so dramatically in recent years that there is little roomfor improvement. However, industry estimates indicate that even smallreductions in the feed conversion ratio measured to the second decimalpoint are significant, and can represent substantial savings to thepoultry producer. Results of the feed conversion analysis are listed inthe table below.

    ______________________________________                                        Results:                                                                      FEED CONVERSION                                                               (Pounds of Feed:Pounds Gained)                                                Group                                                                         Reduction %                                                                            6 Days   13 Days  20 Days                                                                              45 Days                                     ______________________________________                                        Control  0.935    1.232    1.391  1.986                                       Treated  0.922    1.161    1.372  1.975  0.55                                 ______________________________________                                    

Differences in body weight were determined by individual chick weightmeasurement for both control and treated chicks. The table below is anaverage of the body weight data collected on the treated and controlchicks.

    ______________________________________                                        Results:                                                                      BODY WEIGHT                                                                   (Pounds)                                                                      Group                                                                         Reduction %                                                                            6 Days   13 Days  20 Days                                                                              45 Days                                     ______________________________________                                        Control  0.250    0.591    1.324  5.133                                       Treated  0.268    0.667    1.428  5.267  2.61                                 ______________________________________                                    

EXAMPLE 2

Of 1,728 fertilized eggs, 576 were held as controls and 288 weresubjected to a vacuum of 29.8 inches of mercury for ten minutes,pressurized to ambient with filtered air only, and then placed in anincubator as shown in FIG. 2 and FIG. 9.

Another 288 fertilized eggs were placed in a pressure chamber forfifteen minutes with filtered ambient air only at 28 pounds per squareinch and then placed in the same incubator as shown in FIG. 3 and FIG.10.

Another 288 fertilized eggs were subjected to both 29.8 inches ofmercury vacuum for 10 minutes and pressurized with filtered ambient aironly for fifteen minutes at 28 pounds per square inch as shown in FIG. 1(but without U.V. generation of ozone) and FIG. 8.

Another 288 fertilized eggs were subjected to vacuum of 29.8 inches ofmercury for ten minutes and immediately pressurized with O₂ /O₃ forfifteen minutes at 28 pounds per square inch. A mixture of 99.6% pureoxygen was flushed across a 35 watt U.V. (2,537 Å) light until the ozoneconcentration measured by an ozone monitoring device was 4.5 grams percubic meter of O₂. This mixture was flooded into the vacuum chamber torelieve the vacuum over a period of eight minutes and then allowed topressurize the vessel to 28 pounds per square inch over a period offifteen minutes as shown in FIG. 1 and FIG. 8.

All test lots and controls were placed randomly in the same 2,000-eggJamesway incubator (Model No. 1080) at 60% relative humidity and atemperature of 99.5° F.±0.5° F.

    ______________________________________                                        Results:                                                                      Group                                                                         Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Control      488      88        84.7                                          Group #1     235      53        81.6   -3.7                                   (Vacuum only)                                                                 Group #2     238      50        82.6   -2.5                                   (Pressure only)                                                               Group #3     231      57        80.2   -5.3                                   (Pressure/Vacuum)                                                             Group #4     260      28        90.3   +6.6                                   (Vacuum/pressure/                                                             O.sub.3)                                                                      ______________________________________                                    

These results demonstrate the bactericidal effectiveness of pressure andvacuum and the beneficial effects of O₂ /O₃ on fertilized egg mortalitywhen applied prior to incubation and early prenatal phases of the egg.

EXAMPLE 3

Example 1 was repeated, except that no pressure cycle was employed, seeFIG. 1 and FIG. 8, excluding the pressure cycle. The chamber was broughtfrom vacuum to ambient, then exhausted and the eggs were removed forincubation.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     430      90        82.7                                          Group #1     438      82        84.2   1.8                                    Controls     420      100       80.8                                          Group #2     434      86        83.5   3.3                                    Controls     431      89        82.9                                          Group #3     454      66        87.3   5.3                                    ______________________________________                                    

The average increase over control was about 3.5%

This test shows the profound effect of O₂ /O₃ when added to the vacuumintake. (Compare these results with the test results in which O₂ /O₃ wasnot employed.)

EXAMPLE 4

Example 3 was repeated except that only ambient air was drawn into thevacuum intake with no oxygen/ozone, see FIG. 2 and FIG. 9.

Vacuum treatment alone of fertilized eggs without the benefit of oxygen,and particularly ozone, results in significant increased mortality whencompared to the controls.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     433      87        83.3                                          Group #1     418      102       80.4   -3.5                                   Controls     450      70        86.5                                          Group #2     434      86        83.5   -3.5                                   Controls     454      66        87.3                                          Group #3     441      79        84.8   -2.9                                   ______________________________________                                    

EXAMPLE 5

The protocol of Example 3 was followed except that only pressure (nooxygen or ozone) was employed; see FIG. 3 and FIG. 10. Air was suppliedby a two horsepower, two-stroke air compressor through a carbon filter.

Pressure treatment alone of fertilized eggs without benefit of oxygen,and in particular ozone, results in a significant increased mortalitywhen compared to the controls.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     431      89        82.9                                          Group #1     423      97        81.3   -1.9                                   Controls     440      80        84.6                                          Group #2     428      92        82.3   -2.7                                   Controls     456      64        87.7                                          Group #3     441      79        84.8   -3.3                                   ______________________________________                                    

EXAMPLE 6

As shown in FIG. 4 and FIG. 10, pressure treatment alone of fertilizedeggs with oxygen, and in particular ozone, results in significantincreased hatch rates over the controls. The oxygen and ozone weresupplied at thirty two pounds per square inch.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     444      76        85.4                                          Group #1     463      57        89.0   4.2                                    Controls     420      100       80.8                                          Group #2     434      86        83.5   3.3                                    Controls     423      97        81.3                                          Group #3     437      91        84.0   3.3                                    Controls     439      81        84.4                                          Group #4     456      64        87.7   3.9                                    ______________________________________                                    

EXAMPLE 7

As shown in FIG. 1 and FIG. 8 (without pressure cycle), vacuum treatmentalone of fertilized eggs with oxygen, and in particular ozone, resultsin significant increased hatch rates over the controls. Oxygen and ozonewere supplied to 28.5 inches of mercury.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     467      53        89.8                                          Group #1     476      44        91.5   1.9                                    Controls     439      81        84.4                                          Group #2     456      64        87.7   3.9                                    Controls     425      95        81.7                                          Group #3     440      80        84.6   3.5                                    ______________________________________                                    

Chicks from Control 2 and Group 2 were grown separately for twenty-threedays. Mortality in the control group was ten birds, or a loss of 2.3%.For the test group it was seven birds, or a loss of 1.5%.

EXAMPLE 8 TEST 1

Non-invasive injection of hydrogen peroxide solutions through the shellpores of fertile eggs results in significant hatch increase.

500 milliliters of an 8% solution of hydrogen peroxide was added to thereservoir of a 3 milliliter capacity peristaltic delivery pump attachedto a plastic-tipped nozzle five inches long by 1/4 inch in diameter witha bore of approximately one millimeter. The pump line pressure duringthe positive phase measured sixty-five pounds per square inch.

    ______________________________________                                        Results:                                                                      Test 1                                                                        Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     441      79        84.8                                          Group #1     456      64        87.7   3.4                                    Controls     427      93        82.1                                          Group #2     440      80        84.6   3.0                                    Controls     442      78        85.0                                          Group #3     455      65        87.5   2.9                                    Controls     470      50        90.4                                          Group #4     489      31        94.0   4.0                                    ______________________________________                                    

Increased hatch at the use levels of hydrogen peroxide was over 3%.

TEST 2

Referring to FIGS. 5A through 5C, concentrated ozone in air (>1,000parts per million ozone to 99.6% pure oxygen) was directed onto therounded (top) end of fertilized eggs in a high pressure burst. Deliverywas by a controlled delivery, high velocity spring-loaded impact piston(2 cubic centimeters at >100 pounds per square inch).

    ______________________________________                                        Results:                                                                      Test 2                                                                        Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     440      80        84.6                                          Group #1     455      65        87.5   3.4                                    Controls     427      93        82.1                                          Group #2     440      80        84.6   3.0                                    ______________________________________                                    

EXAMPLE 9

Same test as Example 8, except distilled water and 2.0% solution ofpotassium permanganate were used in the injector.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     427      93        82.1                                          Group #1     437      83        84.0   2.3                                    Controls     441      79        84.8                                          Group #2     453      67        87.1   2.7                                    Controls     430      90        82.7                                          Group #3     442      78        85.0   2.8                                    ______________________________________                                    

The increased hatch at use levels of potassium permanganate was about2.5%.

EXAMPLE 10

A Jamesway model 252 incubator 10 was modified in accord with FIG. 6.Specifically, the incubator 10 was reinsulated with 1.5 inch R-90insulation sheets 12 and made air tight by the addition of seals 14 andsilicone caulking along all open seams or leakage points such thatpressures of up to 35 PSI can be attained. Tightly closeable dampers 16and 17 were fixed in the ambient air inlet 18 and outlet 20 lines sothat all incoming or outgoing ambient air could be sealed off by closingor opening the dampers. The condensate exit line 22 was affixed to aknown-in-the-art positive displacement pump (not shown) whichautomatically emptied condensate when humidity increased above 56%+2%for the first eighteen days and 58%+2% for the last three days ofincubation. The condensate pump activated at 58% and 60%, respectively.Oxygen and ozone humidity probe 24 (Cole Parmer humidity/temperaturetransmitter, Model H-37402-00, 4 to 20 ma output, RH 20-95%+2%,temperature 40° to 130° F.), air pressure probe 26 (0 to 100 PSIA, 1 to5 volt, manufactured by Cole Parmer), carbon dioxide probe 28 (ColeParmer CO₂ meter), and temperature probe 30 (type K thermocoupleattached to a strawberry tree analog data acquisition board). Dataacquisition was recorded every ten seconds and stored in process datamonitor 32 (an IBM-compatible computer). Automatic controllers wereoperated for humidity and temperature compensation. Oxygen and ozonewere monitored and physically increased or decreased by opening orclosing an oxygen regulator valve 33 connecting incubator 10 to oxygensource 34, and by activating or deactivating the U.V. ozone generatormodule 36 (2537 Å). CO₂ build-ups were sorbed in an inline impingementscrubber 38 containing 10% KOH and water to limit CO₂ build-up to nomore than 0.5%. Oxygen source 34, ozone generator module 36 and scrubber38 are all connected to air recycle line 40, along with air compressor42 (0 to 250 rpm, 1 cc bursts, 7/16" Tyson Peristaltic variable speedpump). Air recycle line 40, in turn, communicates with recycle inlets 42in the bottom of incubator 10 and recycle outlets 44 in the top ofincubator 10.

The test used one thousand eggs as subjects and one thousand ascontrols. Both the control and the test incubators were modified to thesame specifications except the control incubator was fed from a twohorsepower compressor through an adjustable air regulator. Air intakefrom the compressor was first filtered through a charcoal and fiberfilter. CO₂ was absorbed in the same manner as in the oxygen/ozoneincubator.

The control and the oxygen treatment incubators were set to 99.5°F.±0.5° F. and 60%±2% relative humidity for the first eighteen days and70%±2% relative humidity for the last three days. Gas transport turnoverper incubator was at the rate of about six liters per hour. Thetop-mounted ambient air outlet 20 was adjusted to permit ambient airintake through the partially opened, filtered damper 16 of air inlet 18to achieve an average oxygen concentration of 39% in the oxygenatedincubator 10 with 0.21 grams of ozone per liter of oxygenatedrecirculated air. Inlet 18 communicates with internal fan/heater 45 ofincubator 10 for circulation of heated ambient air therein. Carbondioxide was controlled to a constant <0.5% above ambient levelsthroughout the tests. Pressure was adjusted by opening the damper 16 ofair outlet 20 and damper 17 of inlet 18 until ambient pressure wasachieved. The units were operated immediately after loading for a periodof twenty-eight hours. Immediately afterward the oxygen source 34, U.V.ozone generator module 36 and compressed air source 49 were deactivated.Air intake dampers 17 for both systems were opened and air outletdampers 16 were left in the closed position. Valves 46 of air bleederlines 48 were left about 10% open. Air bleeder line 48 feeds air fromcompressed air source 49 into incubator 10. Eggs are located on racks50. Incubator humidity is controlled by the addition of water throughwater inlet 52 into water collection tray 54 in incubator 10.

Vacuum pump 56 communicates with incubator 10 through line 58 andfacilitates a negative pressure environment within incubator 10. Vacuumpump 56 may be, for example, a Cole Parmer rotary vane high vacuum pump,Model H-74300-65. The vacuum created is preferably between about 2.0inches of mercury and about 29.99 inches of mercury. Note that vacuumpump 56, and at least one of compressed air source 49 and pressurizedoxygen source 34 are employed when sequential application of bothpositive pressure and negative pressure to incubator 10 are desired. If,however, only negative pressure is desired, only vacuum pump 56 isemployed. If only positive pressure is desired, only pressurized oxygensource 34 and/or compressed air source 49 are employed. Air pressure ofup to 35 PSI is attained with the above configuration.

The eggs hatched on the twenty-first day from commencement ofincubation. Oxygen/ozone treated eggs began hatching about eight hoursbefore controls.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     829      171       82.9                                          Group #1     862      138       86.2   4.0                                    Controls     841      159       84.1                                          Group #2     869      131       86.9   3.3                                    Controls     832      168       83.2                                          Group #3     858      142       85.8   3.1                                    ______________________________________                                    

EXAMPLE 11

The protocol of Example 10 was repeated, except that the oxygenatedincubator air pressure was adjusted to five to six pounds per squareinch.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     859      141       85.9                                          Group #1     888      112       88.8   3.4                                    Controls     867      133       86.7                                          Group #2     913       87       91.3   5.3                                    ______________________________________                                    

Chicks from Group 1 and Control 1 were grown separately for three weeks.

Mortality in Group 1 group was ten birds, a loss of 1.2%. In the controlgroup it was seventeen birds, a loss of 2%.

EXAMPLE 12

The protocol of Example 10 was repeated except that the incubators wereoperated normally for forty-eight hours before initiating tests.

    ______________________________________                                        Results:                                                                      Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     831      169       83.1                                          Group #1     858      142       85.8   3.2                                    Controls     800      200       80.0                                          Group #2     834      166       83.4   4.3                                    ______________________________________                                    

EXAMPLE 13

The following examples illustrate the non-invasive addition of chemicalsother than oxygen and ozone through the egg shell without compromisingthe structural integrity of the shell. One thousand fertilized eggs wereheld as controls and one thousand treated according to the protocol ofExample 1.

A 1/2 inch diameter porous bandage was saturated on the adsorption padwith 0.8 milliliter distilled water and 0.3 milliliter of Omaha VaccineCompany, Marek's disease vaccine and affixed to the rounded top (pointof highest porosity) of each of the one thousand eggs to beforce-enriched with oxygen/ozone.

Two hundred fifty control and two hundred fifty oxygen-treated andforced-vaccinated eggs were destroyed after fourteen days and theembryos tested individually for positive and negative vaccine. The otherfive hundred were hatched after twenty-one days from initiation ofincubation.

    ______________________________________                                        Results:                                                                      ______________________________________                                        VACCINE PENETRATION EMBRYO RESULTS                                            Test 1    Positive Negative                                                   ______________________________________                                        Controls   0       250                                                        Treated   247       3                                                         ______________________________________                                        Test 2    Positive Negative                                                   ______________________________________                                        Controls   0       250                                                        Treated   249       1                                                         ______________________________________                                        HATCH RESULTS                                                                 Increase  Hatched  Unhatched   Hatch %                                                                              %                                       ______________________________________                                        Test 1                                                                        Controls  634      116         84.5                                           Treated   659       91         87.9   4.0                                     Test 2                                                                        Controls  629      121         83.9                                           Treated   661       89         88.1   5.0                                     ______________________________________                                    

EXAMPLE 14

Example 13 was repeated using Omaha Vaccine Chic-N-Pox vaccine.

    ______________________________________                                        Results:                                                                      ______________________________________                                        VACCINE PENETRATION EMBRYO RESULTS                                            Test 1    Positive Negative                                                   ______________________________________                                        Controls   0       250                                                        Treated   250       0                                                         ______________________________________                                        Test 2    Positive Negative                                                   ______________________________________                                        Controls   0       250                                                        Treated   246       4                                                         ______________________________________                                        HATCH RESULTS                                                                 Increase  Hatched  Unhatched   Hatch %                                                                              %                                       ______________________________________                                        Test 1                                                                        Controls  642      108         85.6                                           Treated   664       86         88.5   3.4                                     Test 2                                                                        Controls  634      116         84.5                                           Treated   665       85         88.7   5.0                                     ______________________________________                                    

EXAMPLE 15

Marek's vaccine was introduced under the same conditions as Example 6and in the same dosages as in Example 13.

    ______________________________________                                        Results:                                                                      ______________________________________                                        VACCINE PENETRATION EMBRYO RESULTS                                            Test 1    Positive Negative                                                   ______________________________________                                        Controls   0       250                                                        Treated   225       25                                                        ______________________________________                                        HATCH RESULTS                                                                 Increase  Hatched  Unhatched   Hatch %                                                                              %                                       ______________________________________                                        Test 1                                                                        Controls  213       37         85.2                                           Treated   220       30         88.0   3.2                                     ______________________________________                                    

The above results clearly demonstrate the feasibility of obtainingeffective vaccinations through the pores of the fertile egg withoutsustaining losses incurred during the traditional invasive vaccinationsof hatched chicks.

The advantages of vaccinating through the shell is further intensifiedwhen combined with forced enhancement of oxygen methods. Almost any formof vaccination suitable for chicks can be accomplished by methodsdiscussed in the present invention.

EXAMPLE 16

Growth promoters, including antibiotics and micronutrients, are alsoeffectively conveyed non-invasively through the shell withoutcompromising the structural integrity of the shell, thus favorablyinfluencing hatch rate, early chick survival and increased feedconversions. Gallimycin was introduced under same conditions as Example6, using the same dosages as in Example 13.

    ______________________________________                                        Results:                                                                      ______________________________________                                        VACCINE PENETRATION EMBRYO RESULTS                                                      Positive Negative                                                   ______________________________________                                        Controls   0       250                                                        Treated   225       25                                                        ______________________________________                                        HATCH RESULTS                                                                 Increase  Hatched  Unhatched   Hatch %                                                                              %                                       ______________________________________                                        Controls  213       37         85.2                                           Treated   220       30         88.0   3.2                                     ______________________________________                                    

Chlortetracycline, bacitracin, virginiamycin, bambermycin and lincomycinwere all tested on twenty-five eggs by the above procedure. All werefound to have also penetrated the egg to the shell membrane surface.

EXAMPLE 17

Referring to FIGS. 7A through 7H, a suction cup apparatus 100 which isengaged on the shell, preferably at the point of most pores (the egg topabove the air sac), may be used to apply any or all of the foregoingforms of vacuums or pressures in order to non-invasively transferoxygen, ozone and/or other chemicals through the shell. Positivepressure and vacuum lines 102 and 104, respectively, separatelycommunicate with the cup 106 of the cup apparatus 100, as shown in FIGS.7A through 7H. Line 102 applies positive fluid pressure from a sourceknown in the art, and fluid line 104 applies negative fluid pressurefrom a vacuum source known in the art.

Suction cup apparatus 100 has a center plenum 108 for enabling fluidtransfers between the cup 106 and egg. The positive pressure and vacuumlines 102 and 104 communicate through the center plenum 108 of the cupwith on and off valves 110 and 112, respectively, which may be operatedindependently by hydraulic or electronic actuation well known in theart.

FIG. 7A shows the cup apparatus 100 prior to engagement to the shellsurface. FIG. 7B shows the apparatus 100 immediately after engaging thesurface, with negative pressure being applied through line 104. FIG. 7Cshows the egg air sac emptied by negative pressure, with the air of theair sac passing through line 104 of the cup apparatus 100. FIG. 7D showsthe emptied air sac and the cup 106 being pressure-fitted more snugly todeliver the positive pressure fluid treatment through line 102commencing at FIG. 7E. FIG. 7F shows the air sac being filled throughline 102 with treating agent until full at FIG. 7G, whereupon the addedpressure fit is released and the egg disengaged at FIG. 7H.

Using the above apparatus 100, the egg may be engaged by the contactdevice and positive and/or negative pressure applied. If the pressure isnegative, the air sac will be emptied through line 102 until it conformsto the inside of the egg shell. After emptying the air sac, treatingagents such as oxygen and/or ozone, antibiotics, growth promoters,hormones, micronutrients and vaccines may then be introduced underneutral or positive pressure through line 104. If positive pressure isapplied, such as in a fluid stream, then the apparatus can be heldagainst the egg to permit positive pressure to build and force treatingagents through the pores. This action can be continued until the air sachas been refilled with the desired treating agent.

Treatment may be limited to negative pressure, ambient pressure,positive pressure, or combinations thereof. For example, Examples 1, 3,6, 7 and 8 with treating agents selected from any or all of thosedescribed in foregoing examples may be employed with the aboveapparatus.

An advantage to the present treatment is that the egg can be "candied"by illuminating the egg with a light source 114, as shown in FIG. 7D,while being processed in order to view the condition of the air sac.This resulted in the ability to shorten the process times of Example 1as follows: the 28.5 inches of mercury vacuum cycle was run until theair sac conformed to the inside of the egg shell, approximatelytwenty-five seconds, and a hold time at 28.5 inches of mercury ofapproximately forty seconds was used; release of the vacuum with oxygenand/or ozone (time until neutral pressure) was approximately fifteenseconds; the pressure was increased to thirty-two pounds per square inchfor approximately forty-five seconds, until the air sac had returned toat least original size; and release of pressure to ambient wasapproximately fifteen seconds. The total time of the Example 1 protocolwas reduced to approximately two hours and twenty minutes.

Standard incubation protocols of processed eggs is applied to eggs aftertreatment.

    ______________________________________                                        Results:                                                                      Test 1                                                                        Increase     Hatched  Unhatched Hatch %                                                                              %                                      ______________________________________                                        Controls     85       15        85.0                                          Group #1     90       10        90.0   5.9                                    Controls     83       17        83.0                                          Group #2     88       12        88.0   6.0                                    Controls     85       15        85.0                                          Group #3     87       13        87.0   2.4                                    Controls     84       16        84.0                                          Group #4     88       12        88.0   4.8                                    ______________________________________                                    

EXAMPLE 18

Fifteen eggs were each treated by the method of Example 17, wherebyvarious chemicals were non-invasively infused through the egg shell.

    ______________________________________                                        Results:                                                                      Amount                                                                        Amount*                                                                       With O.sub.2 /O.sub.3            Amount*                                      Recovered    Positive   Negative Injected                                     ______________________________________                                        Probiotic mixture                                                                          12         3        100                                          of Strep. lactis and                                                          Strep. faecalis                                                               unknown                                                                       CuCl.sub.2   15         0        15                                           8.00                                                                          KI.sub.2     15         0        1                                            0.65                                                                          KMnO.sub.4   15         0        90                                           60.00                                                                         FeSO.sub.4   15         0        100                                          70.00                                                                         ZnCl.sub.2   15         0        80                                           40.00                                                                         Mo           14         1        1                                            0.80                                                                          Biotin       15         0        1                                            0.20                                                                          Choline      10         5        1,000                                        500.00                                                                        Folic acid   14         1        5                                            0.60                                                                          Niacin       12         3        30                                           12.00                                                                         Pantothenic acid                                                                           15         0        10                                           6.00                                                                          Riboflavin   11         4        5                                            2.00                                                                          Thiamine     10         5        3                                            1.00                                                                          B-6          14         1        3                                            1.00                                                                          B-12 (cobalamin)                                                                           15         0        1                                            0.08                                                                          ______________________________________                                         *Amounts are in milligrams                                               

While particular embodiments of the present invention have beendescribed in some detail herein above, changes and modifications may bemade in the illustrated embodiments without departing from the spirit ofthe invention.

We claim:
 1. An apparatus for non-invasively inducing at least one ofnegative pressure and a fluid having positive pressure into an eggwithout compromising the structural integrity of the egg shellcomprising:a cup contoured to fit an end of an egg, said cup having aninterior surface adapted to contact the egg; a plenum on said cup; apositive pressure line through said plenum and adjacent said interiorsurface of said cup; a negative pressure line through said plenum andadjacent said interior surface of said cup; first valve means in saidpositive pressure line; and second valve means in said negative pressureline such that an egg sac of an egg can be evacuated by opening saidsecond valve means and passing negative pressure through said negativepressure line to said interior surface of said cup while said firstvalve means is closed, and an egg sac can be re-filled with a fluid byopening said first valve means and passing a fluid having positivepressure through said positive pressure line to said interior surface ofsaid cup while said second valve means is closed without compromisingthe structural integrity of the egg shell.
 2. The apparatus of claim 1further comprising a positive pressure source attached to said positivepressure line and a negative pressure source attached to said negativepressure line.
 3. The apparatus of claim 1 further comprising a lightsource oriented to illuminate an egg sufficiently to make visible theair sac of the egg.
 4. The apparatus of claim 1 wherein positivepressure of between about two PSI and about 35 PSI is fed through saidpositive pressure line.
 5. The apparatus of claim 1 wherein negativepressure of between about two inches of mercury and about 29.99 inchesof mercury is fed through said negative pressure line.
 6. The apparatusof claim 1 wherein said fluid is selected from the group consisting ofoxygen, nascent oxygen, liquid oxygen, oxone, ozone, hydrogen peroxide,and potassium permanganate.
 7. An apparatus for non-invasively inducingat least one of negative pressure and a fluid having positive pressureinto an egg without compromising the structural integrity of the eggshell comprising:a cup contoured to fit an end of an egg, said cuphaving an interior surface adapted to contact the egg; a plenum on saidcup; a positive pressure line through said plenum and adjacent saidinterior surface of said cup; a negative pressure line through saidplenum and adjacent said interior surface of said cup; a positivepressure source in said positive pressure line; and a negative pressuresource in said negative pressure line such that an egg sac of an egg canbe evacuated by passing negative pressure from said negative pressuresource through said negative pressure line to said interior surface ofsaid cup, and an egg sac can be re-filled with a fluid by passing afluid having positive pressure from said positive pressure sourcethrough said positive pressure line to said interior surface of said cupwithout compromising the structural integrity of the egg shell.
 8. Theapparatus of claim 7 further comprising first valve means in saidpositive pressure line and second valve means in said negative pressureline.
 9. The apparatus of claim 7 further comprising a light sourceoriented to illuminate an egg sufficiently to make visible the air sacof the egg.
 10. The apparatus of claim 7 wherein positive pressure ofbetween about two PSI and about 35 PSI is fed through said positivepressure line.
 11. The apparatus of claim 7 wherein negative pressure ofbetween about two inches of mercury and about 29.99 inches of mercury isfed through said negative pressure line.
 12. The apparatus of claim 7wherein said fluid is selected from the group consisting of oxygen,nascent oxygen, liquid oxygen, oxone, ozone, hydrogen peroxide, andpotassium permanganate.
 13. An apparatus for non-invasively inducing atleast one of negative pressure and a fluid having positive pressure intoan egg without compromising the structural integrity of the egg shellcomprising:a light source oriented to illuminate an egg sufficiently tomake visible the air sac of the egg; a cup contoured to fit an end of anegg, said cup having an interior surface adapted to contact the egg; aplenum on said cup; a positive pressure line through said plenum andadjacent said interior surface of said cup; a negative pressure linethrough said plenum and adjacent said interior surface of said cup; apositive pressure source in said positive pressure line; a negativepressure source in said negative pressure line; first valve means insaid positive pressure line; and second valve means in said negativepressure line such that an egg sac of an egg can be evacuated by openingsaid second valve means and passing negative pressure from said negativepressure source through said negative pressure line to said interiorsurface of said cup while said first valve means is closed, and an eggsac can be re-filled with a fluid by opening said first valve means andpassing a fluid having positive pressure from said positive pressuresource through said positive pressure line to said interior surface ofsaid cup while said second valve means is closed without compromisingthe structural integrity of the egg shell.
 14. The apparatus of claim 13wherein positive pressure of between about two PSI and about 35 PSI isfed through said positive pressure line.
 15. The apparatus of claim 13wherein negative pressure of between about two inches of mercury andabout 29.99 inches of mercury is fed through said negative pressureline.
 16. The apparatus of claim 13 wherein said fluid is selected fromthe group consisting of oxygen, nascent oxygen, liquid oxygen, oxone,ozone, hydrogen peroxide, and potassium permanganate.